大鼠DRG内ClC-3敲减导致的TNF-α/IL-10失衡对电压门控性钠通道表达和机械痛敏的影响  

Effects of ClC-3 knockdown-induced TNF-α/IL-10 imbalance in rat DRG on voltage-gated sodium channel expression and mechanical allodynia

作　　者：汪小芳 何苏月 马卓琳 杨杰 庞瑞萍 WANG Xiao-fang;HE Su-yue;MA Zhuo-lin;YANG Jie;PANG Rui-ping(Pain Research Center and Department of Physiology,Zhongshan School of Medicine,Sun Yat-sen University,Guangzhou 510080,China;Department of Physiology,School of Medicine,Jinan University,Guangzhou 510632,China)

机构地区：[1]中山大学中山医学院生理学教研室,中山大学疼痛研究中心,广东广州510080 [2]暨南大学基础医学院生理学系,广东广州510632

出　　处：《中国病理生理杂志》2020年第7期1153-1160,共8页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81471132);广东省自然科学基金资助项目(No.2014A030313030,No.2019A1515011295)。

摘　　要：目的:研究敲减大鼠背根神经节(DRG)内ClC-3氯通道/反向转运体对神经元电压门控性钠通道表达和大鼠机械痛敏的影响及机制。方法:成年SD大鼠鞘内注射ClC-3 shRNA腺相关病毒(AAV-ClC-3 shRNA)以敲减DRG内ClC-3的表达,RT-qPCR、免疫荧光染色和Western blot从mRNA和蛋白水平检测ClC-3、细胞因子和电压门控性钠通道表达的变化,用up-down方法检测机械痛敏。结果:鞘内注射AAV-ClC-3 shRNA后DRG组织ClC-3的表达下调,大鼠出现机械触诱发痛。敲减DRG组织ClC-3的表达可致电压门控性钠通道Nav1.3、Nav1.7、Nav1.8和Nav1.9的表达增加。敲减DRG组织ClC-3的表达可提高肿瘤坏死因子α(TNF-α)水平,降低白细胞介素10(IL-10)水平。结论:大鼠DRG内ClC-3下调导致TNF-α/IL-10失衡,进而增加电压门控性钠通道Nav1.3、Nav1.7、Nav1.8和Nav1.9表达,增强神经组织兴奋性,最终导致机械痛敏。AIM:To investigate the effect of ClC-3 chloride channel/antiporter knockdown in rat dorsal root ganglion(DRG)on voltage-gated sodium channel expression in neurons and mechanical allodynia in rats.METHODS:Adeno-associated virus carrying ClC-3 shRNA(AAV-ClC-3 shRNA)was injected intrathecally to knock down ClC-3 expression in DRG tissues of adult SD rats.The mRNA and protein expression levels of ClC-3,cytokines and voltage-gated sodium channels were detected by RT-qPCR,immunofluorescence and Western blot.The mechanical sensitivity was assessed using von Frey hairs and up-down method.RESULTS:Intrathecal injection of AAV-ClC-3 shRNA decreased ClC-3 expression in the DRG tissues and induced mechanical allodynia in the rats.Knockdown of ClC-3 up-regulated the expression levels of Nav1.3,Nav1.7,Nav1.8 and Nav1.9 in the DRG tissues.Knockdown of ClC-3 increased tumor necrosis factor-α(TNF-α)and decreased interleukin-10(IL-10)levels in the DRG tissues.CONCLUSION:Knockdown of ClC-3 in rat DRG tissues induces TNF-α/IL-10 imbalance and increases expression of voltage-gated sodium channels,thus contributing to mechanical allodynia.

关 键 词：神经病理性疼痛 ClC-3通道 背根神经节 肿瘤坏死因子α 白细胞介素10 电压门控性钠通道 机械痛敏 

分 类 号：R741[医药卫生—神经病学与精神病学] R363[医药卫生—临床医学]